External inorganic N source enhances the uptake of As species in garland chrysanthemum (C. coronarium) amended with chicken manure bearing roxarsone and its metabolites.

Roxarsone (ROX), a widely used feed organoarsenic additive, is excreted as itself and its metabolites in animal manure. Animal manure is commonly applied with N fertilizer to meet the N demand of crop. We investigated the accumulation of As species in garland chrysanthemum plants fertilized with chicken manure (CM) bearing ROX and its metabolites, combined with different inorganic N sources (NH4(+), NO3(-) and urea), respectively. The change of pH, N forms and As species in soils was examined as well. The results show that As(V), As(III) and dimethylarsinic acid (DMA) were detectable in soils, and conversions between As species were affected by three inorganic N sources, irrespective of N form and soil pH. As(III) was the sole As species in garland chrysanthemum shoots, and As(III) and As(V) could be detected in roots. Urea, superior to NH4(+), significantly enhanced the uptake of As species in plants by promoting plant growth, while NO3(-) slightly reduced the As accumulation due to decreased biomass. As(III) was the dominant As compound (86.9-89.7%) in plants. Therefore, inorganic N fertilizers may inadvertently increase the risk of As contamination in plant from ROX via the way ROX→chicken→CM→soil→crop.

Roxarsone, inorganic arsenic, and other arsenic species in chicken: a U.S.-based market basket sample.

BACKGROUND: Inorganic arsenic (iAs) causes cancer and possibly other adverse health outcomes. Arsenic-based drugs are permitted in poultry production; however, the contribution of chicken consumption to iAs intake is unknown. OBJECTIVES: We sought to characterize the arsenic species profile in chicken meat and estimate bladder and lung cancer risk associated with consuming chicken produced with arsenic-based drugs. METHODS: Conventional, antibiotic-free, and organic chicken samples were collected from grocery stores in 10 U.S. metropolitan areas from December 2010 through June 2011. We tested 116 raw and 142 cooked chicken samples for total arsenic, and we determined arsenic species in 65 raw and 78 cooked samples that contained total arsenic at ≥ 10 µg/kg dry weight. RESULTS: The geometric mean (GM) of total arsenic in cooked chicken meat samples was 3.0 µg/kg (95% CI: 2.5, 3.6). Among the 78 cooked samples that were speciated, iAs concentrations were higher in conventional samples (GM = 1.8 µg/kg; 95% CI: 1.4, 2.3) than in antibiotic-free (GM = 0.7 µg/kg; 95% CI: 0.5, 1.0) or organic (GM = 0.6 µg/kg; 95% CI: 0.5, 0.8) samples. Roxarsone was detected in 20 of 40 conventional samples, 1 of 13 antibiotic-free samples, and none of the 25 organic samples. iAs concentrations in roxarsone-positive samples (GM = 2.3 µg/kg; 95% CI: 1.7, 3.1) were significantly higher than those in roxarsone-negative samples (GM = 0.8 µg/kg; 95% CI: 0.7, 1.0). Cooking increased iAs and decreased roxarsone concentrations. We estimated that consumers of conventional chicken would ingest an additional 0.11 µg/day iAs (in an 82-g serving) compared with consumers of organic chicken. Assuming lifetime exposure and a proposed cancer slope factor of 25.7 per milligram per kilogram of body weight per day, this increase in arsenic exposure could result in 3.7 additional lifetime bladder and lung cancer cases per 100,000 exposed persons. CONCLUSIONS: Conventional chicken meat had higher iAs concentrations than did conventional antibiotic-free and organic chicken meat samples. Cessation of arsenical drug use could reduce exposure and the burden of arsenic-related disease in chicken consumers.

Development of an ion chromatography-inductively coupled plasma-mass spectrometry method to determine inorganic arsenic in liver from chickens treated with roxarsone.

Roxarsone, (4-hydroxy-3-nitrophenyl)arsonic acid, is an arsenic-containing compound that has been approved as a feed additive for poultry and swine since the 1940s; however, little information is available regarding residual arsenic species present in edible tissues. We developed a novel method for the extraction and quantification of arsenic species in chicken liver. A strongly basic solution solubilized the liver, and ultrafiltration removed macromolecules and particulate material. Ion chromatography separated the species [arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, (4-hydroxy-3-aminophenyl)arsonic acid, (4-hydroxy-3-acetaminophenyl)arsonic acid, and roxarsone] in the extracts, which were then detected by inductively coupled plasma-mass spectrometry. The extraction oxidized most arsenite to arsenate. For fortification concentrations at 2 µg kg(-1) and above, recoveries ranged from 70 to 120%, with relative standard deviations from 7 to 34%. We detected roxarsone, its 3-amino and 3-acetamido metabolites, inorganic arsenic, and additional unknown arsenic species in livers from roxarsone-treated chickens. Both the originating laboratory and a second laboratory validated the method.

Arsenic: a roadblock to potential animal waste management solutions.

The localization and intensification of the poultry industry over the past 50 years have incidentally created a largely ignored environmental management crisis. As a result of these changes in poultry production, concentrated animal feeding operations (CAFOs) produce far more waste than can be managed by land disposal within the regions where it is produced. As a result, alternative waste management practices are currently being implemented, including incineration and pelletization of waste. However, organic arsenicals used in poultry feed are converted to inorganic arsenicals in poultry waste, limiting the feasibility of waste management alternatives. The presence of inorganic arsenic in incinerator ash and pelletized waste sold as fertilizer creates opportunities for population exposures that did not previously exist. The removal of arsenic from animal feed is a critical step toward safe poultry waste management.

The effects of salinomycin and roxarsone on the performance of broilers when included in the feed for four, five, or six weeks and infected with Eimeria species during the starter or grower phase of production.

The timing and magnitude of a coccidian infection, judged by the numbers of oocysts of Eimeria present in the litter, were affected by the duration of medication. In birds medicated for 6 wk and infected at 35 d of age, fewer oocysts were produced than in birds medicated for 4 or 5 wk whether infected at 18 or 35 d of age. Feed conversion at 6, 7, and 8 wk of birds infected at 18 d and medicated for 6 wk was less than that of birds medicated for 4 or 5 wk. Birds infected at 35 d and medicated for 6 wk had a lower feed conversion than birds medicated for 5 wk. Immunity to Eimeria tenella had developed by 8 wk in birds medicated for 4, 5, or 6 wk if infected at 18 d of age. Immunity did not develop in those birds medicated for 6 wk when infected at 35 d.

Use of antibiotics and roxarsone in broiler chickens in the USA: analysis for the years 1995 to 2000.

In 1995, an antibiotic (ANT) was used in starter, grower, and withdrawal (WD) feeds by 94.3, 98.2, and 75.1% of broiler production units, but by 2000, ANT use had declined to 64.8, 66.9, and 48.1% respectively. Roxarsone (ROX) was used in the starter and grower feeds by 69.8 and 73.9% of production units. Bacitracin (BAC) was used more frequently than other antibiotics in the starter and grower feed. Virginiamycin (VIR) was used most frequently in the WD feed. Most units (69.4%) reported use of two different antibiotics. The use of programs comprising two ANT decreased, whereas programs with a single ANT increased during the period of study. A combination of ionophore (ION) + ROX + ANT was employed most frequently in the starter and grower feeds, whereas an ANT alone was used most frequently in the WD ration. The use of ION + ROX + ANT declined from 1995 to 2000, but use of ION + ROX increased. There were no significant differences in calorie conversion whether plants used ION + ROX + ANT, ION + ROX, or ION + ANT. The number of days to rear birds to 2.27 kg was significantly greater for production units using ION + ROX. Mortality was lower for units that used ION + ROX + ANT and ION + ROX than for those that used ION + ANT. Production units that used ION + ANT were more likely to rear birds to a weight greater than 2.5 kg than to 2.0 to 2.5 kg. Units in the South and Central states were more likely to use an ION + ROX than those in the Northeast and Atlantic states, whereas for ION + ROX + ANT the reverse was the case. The cost of medicating with ION + ROX + ANT decreased from 1995 to 1998.

Efficacy evaluation of lasalocid plus roxarsone combination medication with different geographic field strains of Eimeria acervulina.

Performance of broiler chickens medicated with lasalocid alone (at 125 ppm) or in combination with roxarsone (at 50 ppm) was evaluated in battery and floorpen trials after challenge with geographically different field strains of coccidia containing predominately the upper intestinal species Eimeria acervulina. No significant difference in bird performance measured at 6 days postinfection (PI) was observed between lasalocid plus roxarsone-medicated (L+RM) or lasalocid-medicated (LM) birds challenged in separate battery trials with mixed-species inocula from Alabama or Georgia containing 92% or 88% E. acervulina, respectively. In contrast, L+RM birds challenged in another battery trial with a Louisiana mixed-species inoculum containing 92% E. acervulina showed significant reduction in average weight gain at 6 days PI compared with LM-challenged birds. A floorpen trial done with the same Louisiana inoculum showed significant reduction in average bird weight gain at 27 and 35 days of age (6 and 14 days PI) for L+RM-challenged birds compared with both unmedicated-nonchallenged (UMNC) control and LM-challenged birds. The LM+R groups were significantly lower in average bird weight at 27 days of age than the unmedicated-challenged controls. Feed conversions (FCs) for L+RM birds were significantly higher than those for the UMNC control birds during time of challenge (21-27 days of age) and for the 1-to-27-day-of-age time period. No significant difference in FC was seen between the UMNC and LM groups. Results of this study showed that performance of broiler birds medicated with lasalocid plus roxarsone could vary for geographically different mixed-species challenge inocula that contained predominately E. acervulina.

Effects of roxarsone and monensin on digital flexoral tendons of broiler chickens.

Roxarsone and monensin are common poultry feed additives that are used alone or in combination with other drugs to improve growth and feed utilization in young birds. The effects of monensin and roxarsone on the physiology of flexoral tendons of broiler chickens were examined to understand their relationships to leg weakness that have been occasionally associated with these drugs. Day-old chickens were fed either roxarsone or monensin for a period of 6 wk with two regimens of each of the drugs (roxarsone, 45.4 or 90.8 g/ton feed; monensin, 100 or 150 g/ton feed). None of the treatments had any adverse effect on the growth of the birds or caused any significant leg problem. Roxarsone at 45.4 g/ton caused a significant gain in body weight. The biomechanical strength of digital flexoral tendons was measured in several ways. There were no statistical differences in load at break, the modulus of elasticity, or stress or strain levels between different treatment groups and birds that received no medication. There were no differences in collagen, proteoglycan, and pyridinoline content of tendons. Sequential extraction of tendons with different solvents revealed a significant increase in the percentage of guanidine HCl extractible collagens in monensin-treated birds, and a decrease in the acid extractible collagen in both roxarsone- and monensin-treated groups. The relative content of collagen in acid extractible collagens were significantly small relative to total collagen content. Majority of collagen (84 to 90%) was extractible with pepsin. About 8 to 11% of total collagen was resistant to pepsin that was extractible with collagenase; this did not differ between treatment groups. Roxarsone treatment had no effect on the guanidine soluble collagen pool. The effect of monensin on the increase in guanidine soluble pool of collagen may relate to its disruptive effects on cellular secretory processes, which may be of significance in modulating connective tissue function in conjunction with other factors. However, in the present study, neither roxarsone nor monensin alone produced any significant leg problems nor caused any significant differences in the physiology of flexoral tendons or altered their biomechanical properties.

Evaluation of the efficacy of Eimeria maxima oocyst immunization with different strains of day-old broiler and roaster chickens.

The performances of three different strains of male and female broiler or roaster chickens immunized at 1 day of age with a drug-resistant field isolate of Eimeria maxima were evaluated in battery and floorpen immunization trials. No difference in immunization potential was seen between these strains or sexes of day-old chickens immunized with 2500 oocysts/bird of E. maxima and challenged at 10 days of age with 25,000 oocysts/bird of the same isolate in two separate battery trials. All immunized birds showed significant protection for the parameters of weight gain and midintestinal lesion scores at 7 days post challenge (PC) when compared with nonimmunized challenged controls. In two separate floorpen trials, one broiler bird strain (designated strain 2) immunized by the same procedure and challenged at 21 days of age with the equivalent of 25,000 oocysts/bird of the same isolate showed significant protection for weight gain and lesion scores at 7 days PC when compared with nonimmunized controls. Birds in these same floorpen experiments immunized with E. maxima and then fed three different anticoccidial shuttle diets (narasin and nicarbazin 90 grams/ton (g/t) in starter feed and narasin 72 g/t + roxarsone 45.4 g/t in grower feed; nicarbazin 113 g/t in starter feed and narasin 72 g/t + roxarsone 45.4 g/t in grower feed; and monensin 100 g/t in starter and grower feed) showed no interference with elicited immunity with E. maxima challenge for the same parameters measured. No significant difference was seen between any floorpen immunized bird groups for total weight gain at trial terminations (49 days of age).

Toxic responses in F344 rats and B6C3F1 mice given roxarsone in their diets for up to 13 weeks.

Thirteen-week toxicity studies were conducted in groups of 10 F344 rats and B6C3F1 mice of each sex fed roxarsone at 0, 50, 100, 200, 400, or 800 ppm in the diet. Arsenic levels in blood, urine, kidneys, and liver of rats were measured in additional animals of each sex dosed with 100 or 400 ppm roxarsone. Compound-related mortality occurred in both sexes of rats at 800 ppm and mice at 800 and 400 ppm. Significant body weight gain depression occurred in both sexes of rats at 200, 400, and 800 ppm and mice at 800 ppm. Clinical signs of toxicity (trembling, ataxia, and pale skin) were seen primarily in rats and mice at 800 ppm. Lesions associated with roxarsone administration were noted only in the kidney of rats and were characterized by tubular necrosis and mineralization at the corticomedullary junction. Arsenic levels in urine, blood, liver, and kidneys increased over time and were directly proportional to the level of roxarsone in feed. These levels were greater than 6 times higher in rats than in mice and were about 2 time higher in males than in females. The no-observable-effect level for roxarsone toxicity was estimated at 100 ppm for rats and 200 ppm for mice. No hematology or clinical chemistry effects were found in rats or mice of either sex.

Response of broiler chickens to addition of bacitracin methylene disalicylate and roxarsone to diets containing halofuginone.

Studies were conducted at six locations over a 7-yr period to evaluate the response of broiler chickens to bacitracin methylene disalicylate (BMD) and roxarsone in the presence of diets containing 3 ppm halofuginone/kg feed. Treatments consisted of a 2 x 2 factorial arrangement with 0 or 55 mg BMD and 0 or 50 mg roxarsone/kg feed. These additives were fed beginning with day-old chicks and were removed 6 days before termination of the study, which varied in length from 48 to 56 days among locations. Body weights were significantly improved (P less than .05) by the addition of either BMD or roxarsone with a significant interaction (P less than .05) between BMD and roxarsone. Roxarsone improved body weights only in the presence of BMD. Feed utilization was significantly (P less than .05) improved by addition of either BMD or roxarsone, with no interaction between the two products.

Compatibility and anticoccidial activity of lasalocid in combination with roxarsone and antibiotics against Eimeria mixed infection in chicks.

Lasalocid at the concentration of .0075% (68 g/ton) with and without roxarsone 45.4 g/ton was fed in combination with the growth promotants bacitracin methylene disalicylate 200 g/ton, bambermycins 2 g/ton, lincomycin 4 g/ton, nosiheptide 2.5 g/ton, zinc bacitracin 200 g/ton g/ton, and virginiamycin 20 g/ton exhibited a high degree of anticoccidial activity against mixed Eimeria infection in chickens in 9 day challenged battery trials. In these short term challenge trials chicks fed lasalocid, and the lasalocid growth promotant combinations, performed significantly better (P < .05) for growth and anticoccidial efficacy than those fed the growth promotants alone, and the infected, unmedicated controls. In almost all instances, the lasalocid-roxarsone-antibiotic combinations allowed for numerical increases in gains, improvement in feed conversion, and numerical decreases in lesions (in some cases, statistically significant (P < .05) over chicks fed lasalocid alone and/or the lasalocid antibiotic combination. The growth promotants did not interfere with the anticoccidial activity of lasalocid. The growth promotants fed alone exhibited no anticoccidial activity. However, when roxarsone was combined with the antibiotics, the combination resulted in numerically improved performance, reduced mortality, and in most instances, statistically significant decreases in lesions (P .05) over the infected, unmedicated control.

Effects of lasalocid and monensin in combination with roxarsone on lesion reduction and oocyst suppression in chicks infected with Eimeria tenella field isolates.

The anticoccidial activity of lasalocid, monensin, and roxarsone, alone and in combination, was evaluated against eleven Eimeria tenella recent field isolates. Lasalocid was used at 0.0075. 0.01, and 0.0125% activity drug in feed; monensin at 0.0099 and 0.0121%; and roxarsone at 0.005%. Further studies with lasalocid 0.0075%, monensin 0.0099% and roxarsone 0.005 and 0.0025% combinations were carried out against three E. tenella field isolates selected from the aforementioned strains. Lasalocid and monensin each exhibited a high degree of anticoccidial activity at all concentrations tested. Lasalocid and monensin fed in combination with roxarsone showed, in addition to high anticoccidial activity a further reduction in gross lesions and oocysts production, more pronounced at 0.005% level of roxarsone than at 0.0025%, compared to either medication alone or the roxarsone combinations. These positive effects were noted with all strains tested. The practical aspects of these findings are discussed.